title: Design of a large-scale femtoliter droplet array for single-cell analysis of drug-tolerant and drug-resistant bacteria.
abstract: Single-cell analysis is a powerful method to assess the heterogeneity among individual cells, enabling the identification of very rare cells with properties that differ from those of the majority. In this Methods Article, we describe the use of a large-scale femtoliter droplet array to enclose, isolate, and analyze individual bacterial cells. As a first example, we describe the single-cell detection of drug-tolerant persisters of Pseudomonas aeruginosa treated with the antibiotic carbenicillin. As a second example, this method was applied to the single-cell evaluation of drug efflux activity, which causes acquired antibiotic resistance of bacteria. The activity of the MexAB-OprM multidrug efflux pump system from Pseudomonas aeruginosa was expressed in Escherichia coli and the effect of an inhibitor D13-9001 were assessed at the single cell level.
authors.affiliation: Department of Applied Chemistry, Graduate School of Engineering, The University of Tokyo Tokyo, Japan ; Core Research for Evolutional Science and Technology, Japan Science and Technology Agency Tokyo, Japan.
authors.forename: Ryota
authors.initials: R
authors.lastname: Iino
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Matsumoto
authors.forename: Kunihiko
authors.initials: K
authors.lastname: Nishino
authors.forename: Akihito
authors.initials: A
authors.lastname: Yamaguchi
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Noji
keywords: drug efflux; drug resistance; drug tolerance; microdevice; persister; single-cell analysis; transporter